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Production of Antibodies Against Phosphocholine, Phosphatidylcholine, Sphingomyelin, and Lipid A by Injection of Liposomes Containing Lipid A

From the Departments of Pharmacology and Immunology, Walter Reed Army Institute of Research, Washington, D. C. 20012, and the Division of Blood and Blood Products, Bureau of Biologics, Food and Drug Administration, Bethesda, Maryland 20014

Abstract

Liposomes were investigated as a combined carrier and adjuvant to promote the immunogenicity of lipid A derived from endotoxin. Lipid A-containing liposomes, or lipid A alone, were injected once into rabbits either subcutaneously or i.v. Antiserum activity was measured by complement-dependent damage to appropriate indicator liposomes containing trapped glucose. Lipid A in liposomes produced a greater immune response against lipid A than that obtained by injecting identical amounts of lipid A alone, and the response to the liposome-lipid A combination was equivalent to that obtained by a frequently used alternative method, in which the antigen consisted of acid-treated bacterial cells coated with lipid A.

Antiserum, or purified IgG antibodies, from rabbits immunized with liposomes containing lipid A also reacted with liposomes lacking lipid A. The antibodies were inhibited by phosphocholine, and the activity was adsorbed by phosphatidylcholine, or ceramide phosphocholine (sphingomyelin). The anti-phosphocholine antibodies did not cross-react with lipid A, but their binding to liposomes was inhibited by the presence of anti-lipid A antibodies on the surface of the liposomes. An "anti-liposome" response was not observed when either lipid A alone, acid-treated bacterial cells coated with lipid A, or liposomes lacking lipid A were injected.

We conclude that one injection of liposomes containing lipid A produces an immune response against lipid A that is equivalent to that produced by other methods of immunization. Injection of liposomes containing lipid A also produces an immune response against phosphocholine, phosphatidylcholine, and sphingomyelin.

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